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Gold-decorated magnetic nanoparticles modified with hairpin-shaped DNA for fluorometric discrimination of single-base mismatch DNA

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Abstract

The authors describe the use of gold-decorated magnetic nanoparticles (Au/MNPs) in discriminating DNA sequences with a single-base (guanine) mismatch. The Au/MNPs were characterized through dynamic light scattering, X-ray diffraction, superconducting quantum interference device, and UV/visible spectroscopy. They were then conjugated to a probe oligomer consisting of a hairpin-shaped DNA sequence carrying two signalling fluorophores: fluorescein at its 3′ end and pyrene in the loop region. When interacting with the target DNA sequences, the hybridized probe–target duplex renders the pyrene signal (at excitation/emission wavelengths of 345/375 nm) either quenched or unquenched. Quenching (or nonquenching) of the pyrene fluorescence depends on the presence of a guanine (or a nonguanine) nucleotide at the designated polymorphic site. The linear range of hybridization in these Au/MNPs is from 0.1 nM to 1.0 μM of ssDNA. Conceivably, this system may serve as a single-nucleotide polymorphism probe.

Schematic presentation of probe-conjugated Au/MNP preparation (upper panel) and working principle to discriminate DNA with or without single-base (guanine) mismatch sequences at the polymorphic sites (lower panel). Py denotes pyrene-hooked pyrrolocytidine; F denotes fluorescein.

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Acknowledgements

The authors thank the Ministry of Science and Technology, Taiwan (grant nos. MOST 106-2314-B-390-001-MY2, MOST 104-2113-M-002-019-MY2, and MOST 104-2113-M-390-003) and Armed Forces Zuoying General Hospital (grant no. ZBH 105-03) for their support.

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Authors and Affiliations

  1. Department of Materials Science and Engineering, I-Shou University, Kaohsiung, 84001, Taiwan

    Mei-Hwa Lee

  2. Department of Chemical and Materials Engineering, National University of Kaohsiung, Kaohsiung, 81148, Taiwan

    Ching-Chich Leu, Yu-Fan Tseng & Hung-Yin Lin

  3. Division of Pulmonary Medicine, Department of Internal Medicine, Armed-Forces Zuoying General Hospital, Kaohsiung, 81342, Taiwan

    Cheng-Chih Lin

  4. Department of Chemical and Materials Engineering, National University of Kaohsiung, 700, Kaohsiung University Road, Nan-Tzu District, Kaohsiung, 811, Taiwan

    Hung-Yin Lin

  5. Department of Life Sciences, National University of Kaohsiung, Kaohsiung, 81148, Taiwan

    Chia-Ning Yang

  6. Department of Life Sciences, National University of Kaohsiung, 700, Kaohsiung University Road, Nan-Tzu District, Kaohsiung, 811, Taiwan

    Chia-Ning Yang

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  1. Mei-Hwa Lee
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Lee, MH., Leu, CC., Lin, CC. et al. Gold-decorated magnetic nanoparticles modified with hairpin-shaped DNA for fluorometric discrimination of single-base mismatch DNA. Microchim Acta 186, 80 (2019). https://doi.org/10.1007/s00604-018-3192-9

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